LOCUS       sequence-445928-        2518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4G-cloning donor for C-terminal CPD-His10 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2518)
  AUTHORS   Taschner M, Dickinson JB, Roisne-Hamelin F, Gruber S
  TITLE     4G cloning: rapid gene assembly for expression of multisubunit 
            protein complexes in diverse hosts
  JOURNAL   bioRxiv 2024.06.17.599261
REFERENCE   2  (bases 1 to 2518)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.06.17.599261"
COMMENT     Sequence Label: pD(Ct)_CPD-His10
FEATURES             Location/Qualifiers
     source          1..2518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             682..711
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        890..992
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             993..1652
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1059..1078)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2047..2403
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(2456..2489)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gattcgcgac ctactccgga atattaggtc tcaggatctg atgccctcgc gggcggtaaa
       61 atactccata atcaaaatgt taatagctgg ggcccgatta cggttacacc aacgacagat
      121 ggtggtgaaa cccgcttcga cggtcaaatc atcgttcaaa tggaaaacga cccggtagta
      181 gcaaaagcgg cagccaattt agcaggtaaa catgctgaaa gcagtgtggt ggtgcagctc
      241 gattcagacg gcaactatcg cgtggtgtat ggcgatccgt caaaactgga tggaaagcta
      301 cgttggcagt tggtggggca tggtcgcgac cactcagaaa ctaacaatac tcgcttaagt
      361 ggttacagtg ccgatgagtt ggccgtgaaa ttggccaagt tccaacagtc gtttaatcaa
      421 gccgaaaaca tcaacaacaa accggatcac atcagtattg ttggttgttc tttggtgagt
      481 gacgacaagc aaaaaggctt tggtcatcag tttattaacg cgatggatgc gaatggtctt
      541 cgtgtcgatg tctctgttcg tagttctgaa ctggccgtag acgaggcggg acgtaagcat
      601 accaaggacg cgaatggcga ttgggttcaa aaggcagaaa acaacaaagt ttcgctaagc
      661 tgggacgcgc aaggccttga gcaccaccac caccaccacc accaccacca ttaagggaga
      721 cccactgctt gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
      781 ctgctgtcac cgctgagcaa taactatcat aacccctagg tgccatttca ttacctcttt
      841 ctccgcaccc gacatagatc tgggccaact tttggcgaaa atgagacgtt gatcggcacg
      901 taagagggtc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
      961 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     1021 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     1081 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     1141 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     1201 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     1261 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     1321 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     1381 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
     1441 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     1501 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     1561 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     1621 agtactgcga tgagtggcag ggtggggcgt aattttttta aggcagttat tggtgccctt
     1681 aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga
     1741 aagcaaattc gacccggtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta
     1801 ttgctggttt accggtttat tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc
     1861 ctgaggccag tttgctcagg ctctccccgt ggaggtaata attgacgata tgatcattta
     1921 ttctgcctcc cagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     1981 cgtgttccgc ttcctttagc agcccttgcg ccctgagtgc ttgcggcagc gtgaagctaa
     2041 ttcccatgtc agccgttaag tgttcctgtg tcactcaaaa ttgctttgag aggctctaag
     2101 ggcttctcag tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt
     2161 taaaagcctt atatattctt ttttttctta taaaacttaa aaccttagag gctatttaag
     2221 ttgctgattt atattaattt tattgttcaa acatgagagc ttagtacgtg aaacatgaga
     2281 gcttagtacg ttagccatga gagcttagta cgttagccat gagggtttag ttcgttaaac
     2341 atgagagctt agtacgttaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg
     2401 tactatcaac aggttgaact gctgatcaac agatcctcta cgcggccgcg gtaccataac
     2461 ttcgtatagc atacattata cgaagttatc tgccaggcac atgggtttta ctagtatc
//
